Restriction Digest: Partial: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
m (Partial Restriction Digest moved to Restriction Digest: Partial: To consolidate the digests on the menu) |
|||
| (3 intermediate revisions by the same user not shown) | |||
| Line 38: | Line 38: | ||
You can [[Talk:{{PAGENAME}}|discuss this protocol]]. | You can [[Talk:{{PAGENAME}}|discuss this protocol]]. | ||
[[Category:Protocol]] | [[Category:Protocol]] | ||
[[Category:In vitro]] | |||
[[Category:DNA]] | |||
Latest revision as of 05:18, 18 June 2009
Abstract
This protocol can be used to generate genomic DNA fragments of different sizes using a common cutter. This is useful for many screening/probing processes.
Materials
- 10 microcentrifuge tubes
Reagents
- Restriction Endonuclease
- 10X Restriction Endonuclease buffer
- BSA (10 mg/mL)
- Genomic DNA (Concentration > 0.1 μg/μL)
Procedure
- Label the microcentrifuge tubes 1 - 10
- Mix together the following and invert to mix:
- 180μL Genomic DNA
- 20μL 10X Buffer
- 2μL BSA
- Dispense 40μL of this mix into tube 1 and 20μL into tubes 2 - 10 **
From this point keep everything on ice!!! - Add 1μL of restriction-endonuclease to tube 1 and invert to mix.
- Transfer 20μL from tube 1 to tube 2 and invert to mix.
- Repeat the last step transfering 20μL from the preceding tube to the succeeding tube mixing with every transfer.
- Finally remove 20μL from tube 10
- Incubate all 10 tubes for 1 hour at 37°C.
- Finish the digest by submerging all 10 tubes in an 70°C water bath for 20 minutes.
- Run the samples on a gel and choose the size which best fits your application.
Acknowledgments
Acnkowledge any help you had in development, testing, writing this protocol.
References
See OpenWetWare:Biblio for information on how to reference within a wiki.
Specific Protocols
Add links to all the OWW protocols that have been used in making the consensus.
Discussion
You can discuss this protocol.
